Physiological Intracytoplasmic Sperm Injection Research Articles

Sperm selection for ICSI using hylauronan binding in males with high DNA fragmentation does not appear to select against sperm chromosome aneuploidies

Hyalyronic acid (HA) surrounding the human oocyte acts as a natural selector for morphologically normal sperm with hyaluronic acid receptors and minimal DNA fragmentation. An association between high sperm DNA fragmentation and pregnancy loss has been reported, however the clinical value of selecting sperm for intracytoplasmic sperm injection (ICSI) in conjunction with hyaluronic acid binding assays remains to be clarified. The objective of this study was to evaluate the IVF outcomes of sperm with low HA-binding ability (≤65%) following physiological ICSI (PICSI). Retrospective study. Infertility patients with high sperm DNA fragmentation (≥14%; TUNEL assay) that underwent in vitro fertilization (IVF) and PICSI with low HA-binding ability (≤65%; n=23) were evaluated for IVF outcomes including fertilization, blastocyst development and chromosome constitution. Euploid frozen embryo transfer (FET) outcomes which included implantation rate with fetal heart tone, miscarriage rates and live birth rates, were compared to maternally age-matched infertility patients with normal sperm DNA fragmentation that underwent IVF with ICSI. Statistical analysis was performed using Students t-test and Chi square test where appropriate, significance at P<0.05. Mean maternal and paternal ages for these PICSI patients with low HA-binding ability were 37.9±3.7 and 40.9±6.2 years, respectively. High sperm DNA fragmentation (mean 21.5±6.3%) was the only male factor variable significantly associated with these couples. Fertilization and usable blastocyst development with PICSI selected sperm was observed at acceptable rates of 78% and 46%, respectively. Following trophectoderm biopsy, 38% of blastocysts were identified as euploid, significantly less than maternally age-matched ICSI patients at 52% (P<0.05). To date, there have been 12 euploid FETs resulting in comparable good clinical outcomes to maternally age-matched ICSI patients: 66.7% implantation with fetal heart tone, 83.3% live birth and no clinical pregnancy losses. There were 7 cycles that resulted in all aneuploid blastocysts with no FET, thereby calculating a 45.5% live birth rate from oocyte retrieval for PICSI patients with high sperm DNA fragmentation and low HA-binding ability. No significant differences were observed for the chromosomes involved in the errors between the aneuploid blastocysts from the PICSI patients with low HA-binding ability and the maternally age-matched ICSI patients. In conclusion, this study revealed that PICSI patients with high sperm DNA fragmentation and low HA-binding ability have significantly higher blastocyst aneuploidy rates than maternally age-matched ICSI patients. Nevertheless, following a euploid FET these patients exhibited positive clinical outcomes and no pregnancy losses to date.

An efficiency comparison of different in vitro fertilization methods: IVF, ICSI, and PICSI for embryo development to the blastocyst stage from vitrified porcine immature oocytes

BackgroundMost studies carried out to evaluate recovery and development after porcine oocyte vitrification, reported better rates when cryopreserved in embryonic development stages or zygotes, but not in immature oocytes. For this reason, many studies are performed to improve immature oocyte vitrification protocols testing the use of different cryoprotectant concentrations, cooling devices, incubation times; but only a few of them have evaluated which fertilization procedure enhances blastocyst rates in vitrified oocytes. Therefore, this study was aimed to evaluate: 1) if the sperm selection with hyaluronic acid (HA) or polyvinylpyrrolidone (PVP) before injection could play a key role in increasing fertilization and blastocyst formation and 2) the embryo developmental ability and blastocyst production of porcine immature oocytes retrieved after vitrification-warming and co-cultured with granulosa cells during IVM, using different fertilization techniques: in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and conventional ICSI with hyaluronic acid (HA) sperm selection, known as physiological intracytoplasmic sperm injection (PICSI) and.ResultsSperm selected with HA-PICSI displayed a higher percentage of live/acrosome reacted status compared to those in control and exposed to PVP. Higher dead/acrosome reacted rates were obtained after PVP exposure compared to control and HA. In oocytes, viability significantly decreased after IVM in vitrified oocytes. Besides, IVM rates were not different between control denuded oocytes cultured with granulosa cells (DO-GC) and vitrified oocytes. Regarding fertilization parameters, IVF showed higher percentages of total fertilization rate than those obtained by ICSI and PICSI. However, results demonstrate that PICSI fertilization increased the blastocysts formation rate in control DO-GC and vitrified oocytes compared to IVF and ICSI.ConclusionsTo achieve high blastocyst formation rates from vitrified GV oocytes, it is recommended that sperm should be selected with HA instead of PVP before injection since high viability and acrosome reaction rates were obtained. Also, PICSI fertilization was the best method to produce higher blastocyst rates compared to the IVF and ICSI procedures.

Hyaluronic Acid Binding Sperm Selection for assisted reproduction treatment (HABSelect): study protocol for a multicentre randomised controlled trial

IntroductionThe selection of a sperm with good genomic integrity is an important consideration for improving intracytoplasmic sperm injection (ICSI) outcome. Current convention selects sperm by vigour and morphology, but preliminary...

Intervention improves assisted conception intracytoplasmic sperm injection outcomes for patients with high levels of sperm DNA fragmentation: a retrospective analysis.

Sperm DNA fragmentation (SDF) is used in assisted reproductive technology (ART) programs as an indicator for sperm quality, although there is still a lack of consensus as to its clinical utility. In this retrospective study, we examined intracytoplasmic sperm injection (ICSI) outcomes of 1924 infertile patients who underwent SDF analysis using the sperm chromatin integrity test. ART patients were classified as having low [DNA fragmentation index (DFI) <29%] or high SDF (DFI ≥29%) and by whether or not an intervention [physiological intracytoplasmic sperm injection (PICSI), intracytoplasmic morphologically selected sperm injection (IMSI), testicular sperm extraction (TESE)/testicular sperm aspiration (TESA), frequent ejaculation] was performed. High SDF patients who did not have an intervention had a lower fertilization rate and poorer clinical outcomes from blastocyst transfers as compared with low SDF patients; the fertilization rate was 66.0% vs. 70.2% (p=0.042), single embryo transfer (SET) fetal heart pregnancy rate was 28.5% vs. 45.2% (p=0.042), and SET live birth rate was 24.9% vs. 40.6% (p=0.060), respectively. Furthermore, high SDF patients who had an intervention had significantly improved blastocyst transfer outcomes, similar to those of low SDF patients; the SET live birth rate for high SDF intervention patients was 43.8% as compared with 24.9% for high SDF no intervention patients (p=0.037) and 40.6% for low SDF patients (p=0.446). Analysis of the three main intervention subgroups for high SDF patients revealed that TESE/TESA patients had the highest SET live birth rate; in comparison with 24.2% for high SDF patients who did not have an intervention, PICSI patients had 38.3% (p=0.151), IMSI patients had 28.7% (p=0.680), and TESE/TESA patients had 49.8% (p=0.020). Our data suggest that SDF results indicate ICSI outcomes and that patients who have high SDF benefit from an intervention.

Commentary: Physiological Intracytoplasmic Sperm Injection (PICSI), An Alternative to the Standard ICSI Procedure

The rationale of this study is to uncover sperm selection by hyaluronic acid (HA) binding which could advance fertilization pace and embryo quality in Intracytoplasmic sperm injection (ICSI) cycles. HA a sperm binding nonsulfated glycosaminoglycan required as a part of normal fertilisation. Physiological Intracytoplasmic Sperm Injection employs sperm binding to Hyaluronic acid to opt for which sperm to use for Intracytoplasmic sperm injection. PICSI and their latent benefit are still in tranquil being explored. We expect more detailed facts in the coming few years in relation to for whom these modus operandi are most useful apart from cases discussed. Usually if the sperm binding is less than 65% the PICSI dish is used to select the sperm. The medley of sperm selection for ICSI is essential and directly authorise the male input to the embryo therefore visual evaluation alone should certainly not be the only aspect involved when selecting a sperm for ICSI failed cycle twice. But unfit sperm with DNA fragmentation and compromised maturity is able to appear normal and healthy, but if selected for injection/insemination can go ahead to deprived embryo quality and likely a pregnancy loss.

New Trends in Assisted Reproduction Techniques: Cryopreservation, In vitro Fertilization, Intracytoplasmic Sperm Injection and Physiological Intracytoplasmic Sperm Injection

Since that historically the first human birth was registered through in vitro fertilization in 1978, the improvement and creation of new assisted reproduction techniques such as intracytoplasmic sperm injection and physiological intracytoplasmic sperm injection have become of great importance nowadays. Louise Brown was the first baby conceived through in vitro fertilization in England. Since then, several worldwide studies have reported human and domestic animals outcomes. This fact has solved human fertility problems creating new ways of conceiving. Before the success with in vitro fertilization, other assisted reproduction techniques such as cryopreservation were developed. Spermatozoa were the first mammalian cells to be cryopreserved. These events revolutionized all the research in the field of reproductive biology. However, in terms of cryopreservation, intense work needs to be intended since low recovery rates are still reported. The improvement of vitrification-warming procedures is highly important in order to increase embryo development and outcomes. In domestic species, porcine oocytes are the most sensitive cells during vitrification. The future of vitrification success is promising; therefore cryobiologists have a commitment in the field of assisted reproduction. The aim of this review was the acknowledgement of the current status of assisted reproduction techniques, its application, progress and future prospects.

Advanced sperm selection techniques for assisted reproduction.

Assisted reproductive technologies (ART) such as in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) bring together gametes outside of the body to enhance the probability of fertilisation and pregnancy. Advanced sperm selection techniques are increasingly being employed in ART, most commonly in cycles utilising ICSI. Advanced sperm selection techniques are thought to improve the chance that structurally intact and mature sperm with high DNA integrity are selected for fertilisation. Advanced sperm selection strategies include selection according to surface charge; sperm apoptosis; sperm birefringence; ability to bind to hyaluronic acid; and sperm morphology under ultra-high magnification. These techniques theoretically improve ART outcomes. To evaluate the impact of advanced sperm selection techniques on ART outcomes. Systematic search of electronic databases (Cochrane Menstrual Disorders and Subfertility Group Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, PsycINFO, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Latin American and Caribbean Health Science Information Database (LILACS)), trials registers (ClinicalTrials.gov, Current Controlled Trials, World Health Organization International Clinical Trials Registry Platform), conference abstracts (Web of Knowledge) and grey literature (OpenGrey) for relevant randomised controlled trials. We handsearched the reference lists of included studies and similar reviews. The search was conducted in May 2014. We included randomised controlled trials (RCTs) comparing an advanced sperm selection technique versus standard IVF or ICSI or versus another advanced sperm selection technique. We excluded studies of sperm selection using ultra-high magnification (intracytoplasmic morphologically selected sperm injection, or IMSI), as they are the subject of a separate Cochrane review. Quasi-randomised and pseudo-randomised trials were excluded. Our primary outcome measure was live birth rate per woman randomly assigned. Secondary outcome measures included clinical pregnancy per woman randomly assigned, miscarriage per clinical pregnancy and fetal abnormality per clinical pregnancy. Two review authors independently assessed eligibility of studies and risk of bias, and performed data extraction. Disagreements were resolved by consultation with a third review author. Study investigators were consulted to resolve other queries that arose. Risk ratios (RRs) were calculated with 95% confidence intervals (CIs). We planned to combine studies using a fixed-effect model, if sufficient data were available. The quality of the evidence was evaluated using Grades of Recommendation, Assessment, Development and Evaluation (GRADE) methods. Two RCTs were included in the review. Both evaluated sperm selection by hyaluronanic acid binding for ICSI, but only one reported live births. No studies were identified that were related to surface charge selection, sperm apoptosis or sperm birefringence.One RCT compared hyaluronanic acid binding versus conventional ICSI. Live birth was not reported. Evidence was insufficient to show whether there was a difference between groups in clinical pregnancy rates (RR 1.01, 95% CI 0.84 to 1.22, one RCT, 482 women). This evidence was deemed to be of low quality, mainly as the result of poor reporting of methods and findings. Miscarriage data were unclear, and fetal abnormality rates were not reported.The other RCT compared two different hyaluronanic acid binding techniques, SpermSlow and physiological intracytoplasmic sperm injection (PISCI). Evidence was insufficient to indicate whether there was a difference between groups in rates of live birth (RR 1.16, 95% CI 0.65 to 2.05, one RCT, 99 women), clinical pregnancy (RR 1.07, 95% CI 0.67 to 1.71, one RCT, 99 women) or miscarriage (RR 0.76, 95% CI 0.24 to 2.44, one RCT, 41 women). The evidence for these comparisons was deemed to be of low quality, as it was limited by imprecision and poor reporting of study methods. Fetal abnormality rates were not reported. Evidence was insufficient to allow review authors to determine whether sperm selected by hyaluronanic acid binding improve live birth or pregnancy outcomes in ART, and no clear data on adverse effects were available. Evidence was also insufficient to show whether there is a difference in efficacy between the hyaluronic acid binding methods SpermSlow and PICSI. No randomised evidence evaluating sperm selection by sperm apoptosis, sperm birefringence or surface charge was found.Further studies of suitable quality are required to evaluate whether any of these advanced sperm selection techniques can be recommended for use in clinical practice.

A plea for a more physiological ICSI

Intracytoplasmic sperm injection (ICSI) can be considered the most 'revolutionary' in vitro insemination technique because it has efficiently allowed the treatment of male factor infertility. Although ICSI has been successfully and safely applied worldwide for almost 20 years, currently, we have no real knowledge regarding the hypothetical long-term side effects on ICSI adults, given the increased likelihood of spermatozoa with defective nuclear content fertilising the oocytes. The aim of this review article is to investigate the most recent advances of performing ICSI in the safest possible manner, thus, minimising the theoretical hazards of this procedure. To allow for substantiated recommendation which male gametes to choose for physiological ICSI an updated search was performed in Medline and Embase, from 1996 to June 2011. Recent technical advances allow operators to more or less simulate physiological conditions in the laboratory, reducing potential damage to the gametes. It seems possible to prevent fertilisation by DNA-damaged and chromosomal-unbalanced spermatozoa by selecting ICSI sperm by motility and/or maturation markers such as hyaluronic acid or other zona pellucida receptors. Furthermore, novel non-invasive imaging techniques can be valid tools for helping in the morphological selection of ICSI spermatozoa.